LOCUS       sequence-358135-        2815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HiFi-compatible destination vector for secreted overexpression of 
            3C-cleavable, C-terminal 10His-TwinStrep tagged proteins with 
            ExpreS2 Platform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2815)
  AUTHORS   Coker JA, Katis VL, Fairhead M, Schwenzer A, Clemmensen SB, Frandsen
            BU, de Jongh WA, Gileadi O, Burgess-Brown NA, Marsden BD, Midwood 
            KS, Yue WW
  TITLE     FAS2FURIOUS: Moderate-Throughput Secreted Expression of Difficult 
            Recombinant Proteins in Drosophila S2 Cells.
  JOURNAL   Front Bioeng Biotechnol. 2022 May 5;10:871933. doi: 
            10.3389/fbioe.2022.871933. eCollection 2022.
  PUBMED    35600892
REFERENCE   2  (bases 1 to 2815)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Bioeng Biotechnol."; date: "2022-05-5"; volume: "10:871933. doi"; 
            pages: " 10.3389/fbioe.2022.871933. eCollection 2022"
COMMENT     Sequence Label: pExpreS2-1-C3-10H-SIII
FEATURES             Location/Qualifiers
     source          1..2815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          662..794
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     sig_peptide     817..870
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             877..900
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             901..930
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             join(940..963,964..999,1000..1023)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     polyA_signal    1068..1197
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     rep_origin      complement(1332..1920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1412..1431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2177..2195
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2199..2208
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2205..2579
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    2643..2764
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2680..2699)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 atcaatgtat cttatcatgt cttcacgtaa taagtgtgcg gctagcagtc aactactagt
       61 gaatgcccta ctagaagatg tgtgttgcac aaaatgtccc tggaataacc aatttgaagt
      121 gcagatagca gtaaacgtaa gctaatatga atattattta actgtaatgt tttaatatcg
      181 ctggacatta ctaataaacc cactataaac acatgtacat atgtatgttt tggcatacaa
      241 tgagtagttg gggaaaaaat gtgtaaaagc accgtgacca tcacagcata aagataacca
      301 gctgaagtat cgaatatgag taacccccaa attgaatcac atgccgcaac tgataggacc
      361 catggaagta cactcttcat ggcgatatac aagacacaca caagcacgaa cacccagttg
      421 cggaggaaat tctccgtaaa tgaaaaccca atcggcgaac aattcatacc catatatggt
      481 aaaagttttg aacgcgagcg aacttaagag cgccggagta taaatagagg cgcttcgtct
      541 acggagcgac aattcaattc aaacaagcaa agtgaacacg tcgctaagcg aaagctaagc
      601 aaataaacaa gcgcagctga acaagctaaa caatctgcag aagcttggta ccctcgagct
      661 cgtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag
      721 acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc
      781 ctttctctcc acaggaattc tggatccgat ctcaatatga agttatgcat attactggcc
      841 gtcgtggcct ttgttggcct ctcgctcggg agatctttgg aggttctgtt tcagggtcca
      901 caccatcatc accatcacca tcaccaccat ggttctggtt ggagtcaccc acaattcgag
      961 aaaggtggtg gctctggtgg cggttcaggc ggctccgcct ggtcccatcc acagtttgaa
     1021 aagtgaaccg gtcatatgcg gccgcggatc gatcgatatc tgactaaatc ttagtttgta
     1081 ttgtcatgtt ttaatacaat atgttatgtt taaatatgtt tttaataaat tttataaaat
     1141 aatttcaact tttattgtaa caacattgtc catttacaca ctcctttcaa gcgcgtggga
     1201 ctcgatgctc ggcgccactc aaaggcggta atacggttat ccacagaatc aggggataac
     1261 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1321 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1381 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1441 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1501 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1561 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1621 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1681 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1741 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1801 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1861 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1921 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1981 gggattttgc atgcgctaag cgggctttat aaaacgggct gcgggaccag ttttcatatc
     2041 actaccgttt gagttcttgt gctgtgtgga tactcctccc gacaccgaat taattcggat
     2101 ctctgcaagg gattttggtc atgaacaata aaactgtctg cttacataaa cagtaataca
     2161 aggggtgttc atagtataat acgactcact ataggagggc caccatggcc aagttgacca
     2221 gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc
     2281 ggctcgggtt cagccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg
     2341 tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg
     2401 tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact
     2461 tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt
     2521 tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac
     2581 cgacgccgac caacaccgcc ggtccgacgg cggcccacgg gtcccagggg ggtcgacctc
     2641 gaaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2701 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2761 cttatcatgt cttcacgtaa taagtgtgcg gctagcagtc aactagtcca aactc
//
